Allelic allotype inclusion and exclusion among rabbit Ig-bearing lymphocytes of peripheral blood and lymphoid organs

Bessinger, Beverley A.; Molinaro, Giuseppe A.; Teodorescu, Marius; Dray, Sheldon

doi:10.1016/0008-8749(77)90244-1

Cellular Immunology

1977

DOI: 10.1016/0008-8749(77)90244-1

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Allelic allotype inclusion and exclusion among rabbit Ig-bearing lymphocytes of peripheral blood and lymphoid organs

Beverley A. Bessinger¹,

Giuseppe A. Molinaro²,

Marius Teodorescu³

et al.

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Cited by 9 publications

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“…They considered their technique capable of distinguishing endogenous from passively absorbed sIg. Recently Bessinger et al (12) found that 22-31% of sIg + blood lymphocytes of b4b 5 rabbits had both allotypes and that double allotypepositive cells reappeared when pronase-stripped cells were cultured overnight. The proportions of double allotype-positive cells in other tissues was much lower.…”

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Pre-B and B cells in rabbits. Ontogeny and allelic exclusion of kappa light chain genes.

Hayward¹,

Simons²,

Lawton³

et al. 1978

The Journal of Experimental Medicine

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Mononuclear cells containing small amounts of cytoplasmic IgM (clgM+) a but lacking stable surface immunoglobulin (sIg) appear in both mouse and human fetal liver before the earliest sIg + B lymphocytes (1-3). These eIgM+'sIg -cells, called pre-B cells, are the least mature members of the B-cell lineage so far identified in mammals. Well defined allotypic markers on the kappa light chains of rabbit immunoglobulins Co-allotypes) provide a means for the further analysis of pre-B and B-cell development (4). The expression of immunoglobulin molecules by rabbit B cells is controlled by regulatory and/or structural genes for the variable and constant regions of heavy, kappa, and lambda chains occurring as separate clusters at three unlinked, complex loci. The particular immunoglobulin polypeptide chains synthesized by each cell result from the activation of selected members of the linked gene clusters. In heterozygous rabbits, individual plasma cells synthesize Ig of only one of the alternative allotypes (5-6), apparently using genes inherited from one parent but not both. The mechanism of this allelic exclusion has not been established and the stage of B-cell differentiation at which it occurs is uncertain because of the conflicting results of tests for allotype exclusion by B lymphocytes.When blood lymphocytes from b4b 5 rabbits were treated with 125I anti-b4 and antib5 (7), or studied by combined immunofluorescence and radioautography (8), the majority of B lymphocytes had only one sIg allotype detectable but a small percentage *

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mentioning

confidence: 99%

Pre-B and B cells in rabbits. Ontogeny and allelic exclusion of kappa light chain genes.

Hayward¹,

Simons²,

Lawton³

et al. 1978

The Journal of Experimental Medicine

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Expression of b 4 and b 5 ϰ light chain allotypes by B and pre‐B cells in allotype‐suppressed and neutralized b⁴b⁵ rabbits

et al. 1979

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Expression of b 4 and b 5 x light chain allotypes by B and pre-B cells in allotype-suppressed and neutralized b4b5 rabbits*In previous studies, we described a primitive lymphoid cell found in fetal liver and in the bone marrow of older rabbits which contained cytoplasmic IgM but lacked surface IgM detectable by immunofluorescence. In heterozygous b4b5 rabbits, the pre-B cells in which we could detect these kappa chain allotypes appeared to exhibit allelic exclusion. In the present study, we investigated the effects of allotype suppression and its neutralization on the expression of the b 4 and b 5 allotypes by B and pre-B cells from the spleens and bone marrow of b4b5 rabbits. We found that in young allotypesuppressed rabbits, pre-B cells of the suppressed allotype persist in bone marrow when B cells of the suppressed allotype are absent or severely depleted. The persistence of pre-B cells of the suppressed type supports the view that pre-B cells differ in their responsiveness to external influences such as anti-Ig compared to B lymphocytes. Injection of serum with b 5 immunoglobulin into b4b5 animals suppressed 14-23 days previously for b 5 was followed by the appearance of increased proportions of b 5 B cells in spleen within 24 h. Surviving pre-B cells are a likely source of these rapidly appearing B cells as well as of the B cells bearing surface immunoglobulin of the suppressed allotype which appear during the recovery phase of allotype suppression. IntroductionThe b allotypes of rabbit light chains reflect differences in amino acid sequence of the x. chain constant region which are detectable by serological methods. Although there are extensive differences (20-30%) in amino acid sequence between the four b allotypes (b4, b5, b6, and b9) [l-31, they are inherited as simple Mendelian alleles (reviewed in [4, 51). Normally, a heterozygous animal produces ~t chains of both allelic types although an individual cell produces only one of the two alternative forms [6, 71. If a neonatal rabbit is given an injection of antibodies to a single ~t chain allotype, the result will be a long-lasting suppression of the synthesis of that allotype and a compensatory increase in the production of the alternative b allotype in heterozygotes (e.g. b4bS) or of h chains in homozygotes (e.g. b5b5 or b4b4) (reviewed in [8]). This phenomenon, called allotype suppression, has been intensively studied because of the information it may yield concerning mecha-

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Surface immunoglobulin of rabbit peripheral blood lymphocytes: detection of allotypes by fluorescence and rosetting

et al. 1980

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Specific anti-allotype reagents were prepared to detect heavy and light chain allotypes on surface immunoglobulin-positive (sIg+) cells from a3a3b4b4, a2a2b5b5 homozygous and a2a2b4b5 heterozygous rabbits. Total sIg+ peripheral blood lymphocytes (PBL) were detected with fluorescein-labeled goat anti-light chain and sheep anti-rabbit Ig reagents. The total sIg+ cells detected with these reagents and the sum of a and b allotypes individually measured with specific fluorescent anti-allotype reagents (a2 + a3 or b4 + b5) were measured more or ess the same when measured by fluorescence-activated cell sorter (FACS) II flow microfluorometry. The data provided no evidence for single cells expressing both allelic allotypes (allelic inclusion). We found that FACS and resetting techniques were generally equally sensitive. However, we detected a greater proportion of total b4+ plus b5+ cells by rosetting than by fluorescence in some PBL preparations from heterozygous b4b5 rabbits. This was not seen with artificial mixtures of b4b4 and b5b5 cells. The nature of these cells is not yet known. Conceivably, they were not scored as lymphocytes by light-scatter analysis on the FACS and hence were not counted, but were indistinguishable from lymphocytes by microscopy.

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Allelic allotype inclusion and exclusion among rabbit Ig-bearing lymphocytes of peripheral blood and lymphoid organs

Cited by 9 publications

References 22 publications

Pre-B and B cells in rabbits. Ontogeny and allelic exclusion of kappa light chain genes.

Pre-B and B cells in rabbits. Ontogeny and allelic exclusion of kappa light chain genes.

Expression of b 4 and b 5 ϰ light chain allotypes by B and pre‐B cells in allotype‐suppressed and neutralized b⁴b⁵ rabbits

Surface immunoglobulin of rabbit peripheral blood lymphocytes: detection of allotypes by fluorescence and rosetting

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Allelic allotype inclusion and exclusion among rabbit Ig-bearing lymphocytes of peripheral blood and lymphoid organs

Cited by 9 publications

References 22 publications

Pre-B and B cells in rabbits. Ontogeny and allelic exclusion of kappa light chain genes.

Pre-B and B cells in rabbits. Ontogeny and allelic exclusion of kappa light chain genes.

Expression of b 4 and b 5 ϰ light chain allotypes by B and pre‐B cells in allotype‐suppressed and neutralized b4b5 rabbits

Surface immunoglobulin of rabbit peripheral blood lymphocytes: detection of allotypes by fluorescence and rosetting

Contact Info

Product

Resources

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Expression of b 4 and b 5 ϰ light chain allotypes by B and pre‐B cells in allotype‐suppressed and neutralized b⁴b⁵ rabbits